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Overview

  • Product nameAnti-DDDDK tag antibodySee all DDDDK tag primary antibodies ...
  • Description
    Rabbit polyclonal to DDDDK tag
  • Tested applicationsWB, ICC/IF, ICC more details
  • Immunogen

    Peptide xxxDDDDK conjugated to KLH. xxx represents random amino acids. This sequence represents the Enterokinase Cleavage Site.

  • General notesEpitope tags provide a method to localize gene products in a variety of cell types, study the topology of proteins and protein complexes, identify associated proteins, and characterize newly identified, low abundance or poorly immunogenic proteins when protein specific antibodies are not available. Tagging with xxxDDDDK may be done at the N-terminus, N-terminus preceded by a methionine residue, C-terminus, and in internal positions of the target protein. The small size of the epitope tag and its high hydrophilicity tend to decrease the possibility of interference with protein expression, proteolytic maturation, antigenicity and function. The enterokinase cleavage site allows it to be completely removed from the purified fusion proteins.

Properties

  • FormLiquid
  • Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage bufferPBS with 0.1% sodium azide
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • Clonality Polyclonal
  • IsotypeIgG
  • Research Areas

Applications

Our Abpromise guarantee covers the use of ab1162 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB WB: Use at an assay dependent concentration. Antibody was analyzed by western blot using various amino-terminal and carboxy-terminal DDDDK fusion proteins and Invitrogen Positope" R900-40. A 1/5000 dilution gave bands against 0.1, 1.0 and 10 ng of the fusion proteins and 100 ng and 25 ng of the Positope.
ICC/IF ICC/IF: Use at an assay dependent concentration.
ICC ICC: Use at an assay dependent concentration.

Target

  • RelevanceThis is a useful tool for the localisation and characterisation of DDDDK tagged proteins.
  • Alternative names
      DDDDK epitope tag antibodyDYKDDDDK epitope tag antibodyDYKDDDDK tag antibody
      ECS epitope tag antibodyECS tag antibodyEnterokinase Cleavage Site epitope tag antibodyEnterokinase Cleavage Site tag antibodyFLAG antibody
    see all

Anti-DDDDK tag antibody images

  • ab1162 staining DDDDK tag (pink) in neonatal rat cardiomyocytes transduced with Ad-TBX18/FLAG-IRES-GFP by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and incubated with primary antibody. An AlexaFluor®-conjugated anti-rabbit IgG was used as the secondary antibody. Nuclei were stained with DAPI (blue).
  • ab1162 staining DDDDK tag in hTERT-RPE1 by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with Paraformaldehyde, permeabilized with Saponin and blocked with 0.02% BSA for 20 minutes at 18°C. Samples were incubated with primary antibody (1/100) for 45 minutes at 18°C. A Cy3®-conjugated Donkey anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

References for Anti-DDDDK tag antibody (ab1162)

This product has been referenced in:
  • Arboleda VA  et al. Mutations in the PCNA-binding domain of CDKN1C cause IMAGe syndrome. Nat Genet 44:788-92 (2012). Read more (PubMed: 22634751) »
  • Pillai VB  et al. Acetylation of a conserved lysine residue in the ATP binding pocket of p38 augments its kinase activity during hypertrophy of cardiomyocytes. Mol Cell Biol : (2011). Read more (PubMed: 21444723) »

See all 12 Publications for this product

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (hTERT-RPE1)
Specification hTERT-RPE1
Fixative Paraformaldehyde
Permeabilization Yes - saponin
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 0.02% · Temperature: 18°C
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Submitted Dec 19 2012

Application Western blot
Sample Human Cell lysate - whole cell (hTERT-RPE1)
Loading amount 50000 cells
Specification hTERT-RPE1
Treatment transfected with flag tag plasmid 20hr
Gel Running Conditions Reduced Denaturing (8 % acrylamid)
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Dec 14 2012

Application Western blot
Sample Human Cell lysate - whole cell (HeLA)
Loading amount 25 µg
Specification HeLA
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Submitted Jan 21 2012

Application Western blot
Sample Mouse Recombinant protein (Flag-tagged recombinant protein)
Loading amount 2 µg
Specification Flag-tagged recombinant protein
Gel Running Conditions Reduced Denaturing (4-20% Tris-Glycine Gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Aug 06 2010

Application Western blot
Sample Human Cell lysate - whole cell (Transfected HT1080 Cells)
Loading amount 40000 cells
Specification Transfected HT1080 Cells
Gel Running Conditions Reduced Denaturing (10% SDS-PAGE)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Feb 01 2010

Application Immunocytochemistry
Sample African Green Monkey Cultured Cells (Cos7 cells)
Specification Cos7 cells
Fixative Paraformaldehyde
Permeabilization Yes - 10 min in 0.1% PBS-tween20
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
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Submitted Apr 16 2009

Application Western blot
Sample Rabbit Cell lysate - whole cell (HCT116)
Loading amount 30 µg
Specification HCT116
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Dec 21 2007

Application Immunocytochemistry/ Immunofluorescence
Sample Rabbit Cell (HCT116)
Specification HCT116
Fixative Formaldehyde
Permeabilization Yes - 0.2% Triton-X 100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Submitted Oct 09 2007

Application Western blot
Sample Human Cell lysate - whole cell (Mono Mac 1)
Loading amount 50 µg
Specification Mono Mac 1
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5
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Submitted Mar 20 2006

Application Western blot
Sample Human Cell lysate - whole cell (HEK 293)
Specification HEK 293
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5%
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Submitted Jan 05 2006

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"